Proliferative Phase Endometrium Research Articles

Endometrial Proliferative Phase-Centered View of Transcriptome Dynamics across the Menstrual Cycle.

The endometrium, the inner mucosal lining of the uterus, undergoes complex molecular and cellular changes across the menstrual cycle in preparation for embryo implantation. Transcriptome-wide analyses have mainly been utilized to study endometrial receptivity, the prerequisite for successful implantation, with most studies, so far, comparing the endometrial transcriptomes between (i) secretory and proliferative endometrium or (ii) mid-secretory and early secretory endometrium. In the current study, we provide a complete transcriptome description of the endometrium across the entire menstrual cycle and, for the first time, comprehensively characterize the proliferative phase of the endometrium. Our temporal transcriptome analysis includes five time points including the mid-proliferative, late proliferative (peri-ovulatory phase), early secretory, mid-secretory, and late secretory phases. Thus, we unveil exhaustively the transitions between the consecutive proliferative and secretory phases, highlighting their unique gene expression profiles and possible distinct biological functions. The transcriptome analysis reveals many differentially expressed genes (DEGs) across the menstrual cycle, most of which are phase-specific. As an example of coordinated gene activity, the expression profile of histone-encoding genes within the HIST cluster on chromosome 6 shows an increase in cluster activity during the late proliferative and a decline during the mid-secretory phase. Moreover, numerous DEGs are shared among all phases. In conclusion, in the current study, we delineate the endometrial proliferative phase-centered view of transcriptome dynamics across the menstrual cycle. Our data analysis highlights significant transcriptomic and functional changes occurring during the late proliferative phase-an essential transition point from the proliferative phase to the secretory phase. Future studies should explore how the biology of the late proliferative phase endometrium impacts the achievement of mid-secretory endometrial receptivity or contributes to molecular aberrations leading to embryo implantation failure.

AI-algorithm training and validation for identification of endometrial CD138+ cells in infertility-associated conditions; polycystic ovary syndrome (PCOS) and recurrent implantation failure (RIF)

BackgroundEndometrial CD138+ plasma cells serve as a diagnostic biomarker for endometrial inflammation, and their elevated occurrence correlates positively with adverse pregnancy outcomes. Infertility-related conditions like polycystic ovary syndrome (PCOS) and recurrent implantation failure (RIF) are closely associated with systemic and local chronic inflammatory status, wherein endometrial CD138+ plasma cell accumulation could also contribute to endometrial pathology. Current methods for quantifying CD138+ cells typically involve laborious and time-consuming microscopic assessments of only a few random areas from a slide. These methods have limitations in accurately representing the entire slide and are susceptible to significant biases arising from intra- and interobserver variations. Implementing artificial intelligence (AI) for CD138+ cell identification could enhance the accuracy, reproducibility, and reliability of analysis. MethodsHere, an AI algorithm was developed to identify CD138+ plasma cells within endometrial tissue. The AI model comprised two layers of convolutional neural networks (CNNs). CNN1 was trained to segment epithelium and stroma across 28,363 mm2 (2.56 mm2 of epithelium and 24.87 mm2 of stroma), while CNN2 was trained to distinguish stromal cells based on CD138 staining, encompassing 7345 object layers (6942 CD138- cells and 403 CD138+ cells). The training and performance of the AI model were validated by three experienced pathologists. We collected 193 endometrial tissues from healthy controls (n = 73), women with PCOS (n = 91), and RIF patients (n = 29) and compared the CD138+ cell percentages based on cycle phases, ovulation status, and endometrial receptivity utilizing the AI model. ResultsThe AI algorithm consistently and reliably distinguished between CD138- and CD138+ cells, with total error rates of 6.32% and 3.23%, respectively. During the training validation, there was a complete agreement between the decisions made by the pathologists and the AI algorithm, while the performance validation demonstrated excellent accuracy between the AI and human evaluation methods (intraclass correlation; 0.76, 95% confidence intervals; 0.36–0.93, p = 0.002) and a positive correlation (Spearman's rank correlation coefficient: 0.79, p < 0.01). In the AI analysis, the AI model revealed higher CD138+ cell percentages in the proliferative phase (PE) endometrium compared to the secretory phase or anovulatory PCOS endometrium, irrespective of PCOS diagnosis. Interestingly, CD138+ percentages differed according to PCOS phenotype in the PE (p = 0.03). On the other hand, the receptivity status had no impact on the cell percentages in RIF samples. ConclusionOur findings emphasize the potential and accuracy of the AI algorithm in detecting endometrial CD138+ plasma cells, offering distinct advantages over manual inspection, such as rapid analysis of whole slide images, reduction of intra- and interobserver variations, sparing the valuable time of trained specialists, and consistent productivity. This supports the application of AI technology to help clinical decision-making, for example, in understanding endometrial cycle phase-related dynamics, as well as different reproductive disorders.

Decreased NSD2 impairs stromal cell proliferation in human endometrium via reprogramming H3K36me2.

The proliferation of the endometrium is regulated by histone methylation. This study shows that decreased NSD2 impairs proliferative-phase endometrial stromal cell proliferation in patients with recurrent implantation failure via epigenetic reprogramming of H3K36me2 methylation on the promoter region of MCM7. Recurrent implantation failure (RIF) is a formidable challenge in assisted reproductive technology because of its unclear molecular mechanism. Impaired human endometrial stromal cell (HESC) proliferation disrupts the rhythm of the menstrual cycle, resulting in devastating disorders between the embryo and the endometrium. The molecular function of histone methylation enzymes in modulating HESC proliferation remains largely uncharacterized. Herein, we found that the levels of histone methyltransferase nuclear receptor binding SET domain protein 2 (NSD2) and the dimethylation of lysine 36 on histone H3 are decreased significantly in the proliferative-phase endometrium of patients with RIF. Knockdown of NSD2 in an HESC cell line markedly impaired cell proliferation and globally reduced H3K36me2 binding to chromatin, leading to altered expression of many genes. Transcriptomic analyses revealed that cell cycle-related gene sets were downregulated in the endometrium of patients with RIF and in NSD2‑knockdown HESCs. Furthermore, RNA-sequencing and CUT&Tag sequencing analysis suggested that NSD2 knockdown reduced the binding of H3K36me2 to the promoter region of cell cycle marker gene MCM7 (encoding minichromosome maintenance complex component 7) and downregulated its expression. The interaction of H3K36me2 with the MCM7 promoter was verified using chromatin immunoprecipitation-quantitative real-time PCR. Our results demonstrated a unifying epigenome-scale mechanism by which decreased NSD2 impairs endometrial stromal cell proliferation in the proliferative-phase endometrium of patients with RIF.

Histopathological Analysis of Hysterectomy Specimens in a Tertiary Care Centre: A Retrospective Study.

Introduction The uterus is a crucial reproductive organ that is susceptible to the development of several non-neoplastic and neoplastic diseases in women, greatly increasing morbidity and mortality. Although there are various therapeutic options, hysterectomy is still a popular treatment option throughout the world.Abnormal uterine bleeding, pelvic pain, pelvic inflammatory disease (PID), prolapse of the uterus, adenomyosis, endometriosis, fibroids, gynecological malignancies, and obstetric problems that require hysterectomy, all samples must be examined histopathologically.Histopathological examination of the specimens obtained after hysterectomy is important for both diagnosis and treatment. The current work aimed to identify the various clinical indications, analyze the clinicopathological correlation in hysterectomy specimens, and analyze the patterns of lesions in hysterectomy specimens. Materials and methods This study was conducted in the Department of Pathology at the Datta Meghe Medical College, Wanadongari, Nagpur, from February 2022 to January 2023. All types of hysterectomy specimens received during this year were examined, and the tissues were processed and stained with H&E. Histopathological examination was performed, and various lesions in the hysterectomy specimens were examined.The study included all forms of hysterectomy, including abdominal, vaginal, laparoscopic, and total abdominal hysterectomy. Results An analysis of 110 cases of hysterectomy revealed that abdominal hysterectomy was the type of hysterectomy in 79 (71.82%) cases, with a maximum age range of 35 to 45 years (42.72%). The proliferative phase endometrium was the most common endometrial pathology, accounting for 43 (39.09%) cases, followed by the atrophic endometrium in 35 (31.82%) cases. Leiomyoma was the most prevalent myometrial lesion, accounting for 52 (47.28%) cases, followed by adenomyosis, accounting for 23 (20.91%) cases. Chronic cervicitis was the most common incidental finding in the hysterectomy samples, accounting for 85 (77.28%) cases. Follicular cysts, representing 22 (20%) cases, were the most common ovarian lesions, followed by serous cystadenoma in seven (6.37%) cases. Two cases of malignant tumors were noted: one case of endometrial carcinoma and one case of mucinous cystadenocarcinoma of the ovary. In most cases, ranging from 70% to 100%, the final histopathological diagnosis supports the preoperative clinical diagnosis. Conclusion Hysterectomy is the most common major gynecological surgery performed under elective conditions. Although histological studies and clinical diagnoses are closely correlated, several lesions, including chronic cervicitis and adenomyosis, were discovered incidentally. Therefore, every hysterectomy specimen must undergo a thorough histological investigation, even if it appears superficially normal, to confirm the diagnosis and improve postoperative care.

P-346 Migration-associated microRNAs are dysregulated in endometriosis: potential diagnostic biomarkers

Abstract Study question What is the role of dysregulated endometrial microRNAs (miRNAs) in the development of endometriosis? Summary answer Dysregulated miRNAs in the endometrium of women with endometriosis can also be found in endometriomas and may affect the migratory ability of endometriotic cells. What is known already The molecular mechanisms underlying the pathogenesis of endometriosis are poorly understood. One of the hypotheses is that changes in cell properties observed in endometriotic lesions could be initiated already in the endometrium. Dysregulation of miRNAs in the endometrium of women with endometriosis has been reported in previous microarray-based studies. However, little overlap has been seen between published miRNA expression data. Moreover, the potential role of dysregulated miRNAs in the endometrium of women with endometriosis and whether these changes are also present in the endometrioma are largely unknown. Study design, size, duration In this experimental case-control study miRNA gene expression in proliferative phase endometrium was compared between 15 women with laparoscopically confirmed endometriosis (stage III–IV) and 17 age-matched controls who were laparoscopically confirmed to be free of endometriosis. Selected miRNAs were further compared between paired proliferative-phase endometrium and endometrioma from a new cohort of women with endometriosis (stage III–IV) and studied in vitro to understand their effect on cell migration. Participants/materials, setting, methods Samples were collected during laparoscopic operations performed at Tartu University Hospital, Estonia. Dysregulated endometrial miRNAs were detected with small RNA sequencing and differential gene expression analysis. Target genes and biological pathways were predicted to understand their potential role in the disease. Selected miRNAs were further studied for their expression in endometriomas using qRT-PCR and in-vitro for their proliferation and migration ability using a transwell-migration assay after miRNA mimic transfection of the 12Z endometriotic cell line. Main results and the role of chance In total, we identified 9 upregulated and 5 downregulated (-2 &amp;lt;fold change &amp;gt; 2, FDR &amp;lt; 0.05) miRNAs in the endometrium of women with endometriosis compared to controls. In-silico analyses showed that predicted target genes of the dysregulated miRNAs were significantly enriched in migration-related KEGG pathways such as adherens junctions, focal adhesion, MAPK-, PI3-AKT-, and TGF-beta signaling. The most down-regulated miRNA, miR-193b-5p, and the most up-regulated miRNA, miR-374b-5p, were selected for further characterization. Validation of their expression in endometriomas showed a significant up-regulation of both miRNAs compared to paired endometrium (Fold change &amp;gt; 2, FDR &amp;lt; 0.05). Since it has been reported that altered cell migratory ability could be involved in the pathogenesis of endometriosis and our dysregulated miRNA were associated with migration-related pathways, we explored if miR-193b-5p mimic transfection affects the migration capacity of 12Z cells. A 2-fold decrease in cell migration (p-value 0.0021) was observed after 12Z cell mimic transfection. No change in proliferation was demonstrated. Limitations, reasons for caution Although our findings both in-silico and in-vitro suggest a link between dysregulated miRNAs and cell migration, further in-vitro studies in primary cells and in-vivo studies in animal models are needed to reveal the specific pathways that these miRNAs regulate to explain the observed functional changes in the context of endometriosis. Wider implications of the findings This study gives molecular insight into the pathogenesis of endometriosis, a poorly understood disease, by demonstrating changes in miRNA expression in both the endometrium and endometrioma that potentially can be linked to a changed cell migratory ability. Furthermore, identified miRNAs could be further evaluated as diagnostic biomarkers in larger studies. Trial registration number Not applicable

P-625 Artificial intelligence-enabled analysis of endometrial CD138 positive plasma cells in infertility-associated conditions; Polycystic ovary syndrome (PCOS) and recurrent implantation failure (RIF)

Abstract Study question Can artificial intelligence (AI) algorithm identify CD138+ cells in the endometrial stroma, and is the expression altered in PCOS and RIF? Summary answer CD138+ cell detection by AI shows high accuracy. The cell count in controls was comparable to that in PCOS cases but differed in RIF patients. What is known already Immunohistochemical (IHC) staining for CD138+ plasma cells has been applied to detect endometrial inflammation; however, due to a lack of cost-effective and reliable tools, there is no golden standard for CD138+ analysis. PCOS is a common endocrine disorder characterized by irregular menstrual cycles and hyperandrogenism. The women are also affected by systemic low-grade inflammation, and the PCOS endometrium shows dysregulated immune profile. RIF is one of the main causes of a low IVF success rate, and chronic endometrial inflammation has been detected in women with RIF. To date, AI technology has been applied in the diagnostics of various clinical conditions. Study design, size, duration In this study, AI analysis was done for CD138+ cells in a total of 193 endometrial biopsy samples: 73 were obtained from healthy and fertile controls, 91 from women with PCOS (ovulatory samples; n = 78, anovulatory samples; n = 13, Rotterdam criteria), and 29 samples from RIF patients on day five after the initiation of P4 administration. Participants/materials, setting, methods The biopsy samples were collected by suction curette (Pipelle), and endometrial receptivity was assessed by gene expression profiling (beREADY test). The tissues were stained with CD138 antibody, and the AI algorithm, AITAH, was trained with 150 whole slide images (WSIs) to segment epithelium and stroma and segregate CD138- and CD138+ cells in the stroma. AITAH was validated by three external pathologists. The cell counts were compared according to cycle phases, ovulatory status, and endometrial receptivity. Main results and the role of chance The performance of AITHA was excellent, as the final training error for CD138+ cells was 3.23%, and the decisions for the cells between pathologists and AITAH were in complete agreement. The CD138+ cell percentages were higher in the proliferative phase (PE) endometrial tissue than in the ones obtained in the secretory phase (PCOS, p &amp;lt;0.001 or control, p &amp;lt;0.001), regardless of PCOS diagnosis. Similar CD138+ cell percentages were observed between PE endometrium and anovulatory endometrium in PCOS cases (p &amp;lt;0.001). The count of CD138+ plasma cells was negatively correlated with the endometrial thickness (PCOS, p =0.001 or control, p =0.009) and P4 (PCOS, p =0.001 or control, p =0.003), and positive correlations between the cell counts and gonadotropin levels were observed in ovulatory PCOS samples (FSH, p =0.001 and LH, p =0.031). More CD138+ cells were seen in RIF patients compared to the controls at the receptive stage (p = 0.046); however, there was no difference between RIF samples of three different receptivity statuses (p =0.426). Limitations, reasons for caution Further validation for AITAH using confirmed chronic endometritis cases as positive controls is required. And further analysis using control samples obtained during the hormone replacement treatment is required. The small number of anovulatory PCOS subjects may explain the absence of correlations between the CD138+ plasma cell counts and clinical characteristics. Wider implications of the findings AI-enabled analysis tools could improve histological examination of endometrial immune cells by analyzing high amounts of whole tissue slides within a short time with a high pixel resolution, thus overcoming many current impediments in manual microscopic evaluation. Trial registration number NA

Histopathological Diagnosis of Hysterectomy Specimens in Abnormal Uterine Bleeding

Background: Abnormal uterine bleeding (AUB) is a common and distressing condition effecting predominantly perimenopausal women. Heavy menstrual bleeding (HMB) is the most common presentation of AUB. There are many structural and non structural causes of AUB. Histopathology of endometrial curettings is well studies before but the histopathological patterns of hysterectomy specimens are not well studied. Objective: The objective of this study is to: Find out histopathological diagnosis of hysterectomy specimens in patients of AUB. Study Design: Retrospective Cohort study. Place and Duration of Study: Gynecology and Pathology departments of Azra Naheed and Rashid Latif medical colleges from January 2022 to January 2023. Methods: Histopathology reports of all hysterectomies done for AUB in both medical colleges allied hospitals were retrieved retrospectively and evaluated in detail. Four age groups were defined 30-40 years, 41-50, 51-60 and 61-70 years. Various patterns were recorded overall and separately for each group and results established. Results: A total of 221 abdominal hysterectomy histopathology reports were retrieved and evaluated. All hysterectomies were performed for AUB during one year period between January 2022 to January 2023. 40(18.09%) women were between 30-40 years of age, 134(60.6%) were between 41-50 years of age, 39(17.6%) were between 51-60 years of age and 08(3.61%) patients were between 61-70 years of age. Distorted proliferative phase endometrium was seen in 28(12.6%) of cases, Adenomyosis was seen in 76(34.38%), endometrial polyps in 19(8.55%), fibroids in 92(41.6%), metaplasia 07(3.1%), hyperplasia 10(4.5%), and endometrial carcinoma was seen 09 (4.07%) of cases. Conclusion: AUB is most common in perimenopausal women. Fibroids, Adenomyosis, and disordered proliferative endometrium were most common in perimenopausal women while carcinoma endometrium was most common cause of AUB in postmenopausal women. This study will help Gynecologists to better find out the cause of AUB and treat accordingly. Keywords: Abnormal uterine bleeding (AUB), Heavy menstrual bleeding (HMB), Perimenopausal, Hysterectomy. Histopathology, Endometrium

Adverse impact of CD138+ cells in proliferative-phase endometrium on pregnancy outcomes in fresh IVF/ICSI cycles.

To evaluate the impact of Syndecan-1 (CD138) in proliferative-phase endometrium on pregnancy outcomes in fresh in vitro fertilization (IVF)/ intracytoplasmic sperm injection (ICSI) cycles. This retrospective cohort study contained 273 patients who underwent IVF/ICSI with fresh embryo transfer following an endometrial curettage from January 2020 to May 2022. Endometrial curettage was performed on all patients within 3 to 5 days following menstruation and endometrial tissue was acquired for detection of plasma cells by immunohistochemistry. Subsequent pregnancy outcomes of all cycles were traced and analyzed. A total of 149 patients became pregnant (i.e., pregnant group) in the fresh transfer IVF/ICSI cycles and 124 did not become pregnant (i.e., nonpregnant group). The number of CD138 + cells/ high-power field (HPF) of the nonpregnant group was significantly higher than the pregnant group (2.36 ± 4.24 vs 1.31 ± 3.41, P = .008). The cut off value of CD138 + cells/HPF was 2 by receiver operating characteristic curve analysis, with an area under the receiver operating characteristic curve of 0.572. Compared with the negative group (i.e., CD138 + cells/HPF < 2, n = 204), the positive group (i.e., CD138 + cells/HPF ≥ 2, n = 69) had a significantly lower clinical pregnancy rate (71.8% vs 40.6%, P < .001). The clinical pregnancy rate revealed a gradually decreasing trend with the increase in CD138 + cells. Proliferative-phase endometrial CD138 + cells may be an adverse indicator for pregnancy outcomes in fresh IVF/ICSI cycles, with a certain value in predicting non-pregnancy. Pregnancy outcome was poor when CD138 + cells/HPF ≥ 2 in the endometrium and may worsen with the increase in CD138 + cells.

HISTOPATHOLOGICAL SPECTRUM OF ENDOMETRIAL LESIONS IN TERTIARY CARE CENTRE- A PROSPECTIVE STUDY

Introduction: Endometrial sampling is most often performed for abnormal uterine bleeding. Irregular cycles, abnormal and dysfunctional uterine bleeding is considered to be one of the most common presenting symptom in gynaecological practice amongst all age group of patients. Early detection of precursor lesions and exclusions of malignancy is a diagnostic challenge for pathologist. This study aims at evaluation of the histopathological spectrum of endometrial lesions common for particular age group, presenting complaints and underlying cause for the same. Materials and methods: A prospective study for a period of 2 years from January 2021 to December 2022 in the Department of Pathology. Total of 870cases , between 18-80years were included. Histopathological study was done and each sample was categorized on the basis of age, bleeding pattern and histomorphological patterns. The mean age was found out to be 42.5±2 years. M Results: ost common age group was 3rd and 4th decade with 231 patients (26.55%) of 31-40 years and 226 patients (25.97%) of 41-50 years with lower abdominal pain and menorrhagia [210 cases (24.13%) &amp; 191cases(21.83%) respectively], proliferative phase endometrium was noted in 35.74% cases, disordered proliferative endometrium and endometrial polyps in 10.34% cases , non atypical hyperplasia was seen in 8.39% cases, pill endometrium in 3.33% and endometrial carcinoma contributed to 0.46%. Perimenopausal period is a vulnerable age group to a numbe Conclusion: r of endometrial lesions. Lower abdominal pain and menorrhagia were most common clinical presentation and confronted wide spectrum of histomorphological changes in endometrial samples presenting with various patterns of bleeding ranging from normal cyclical changes to malignancy. It is observed that patients with obesity, diabetes mellitus, sedentary lifestyle and low socioeconomic status are more proned to these conditions. Histopathological examination plays a vital role in diagnostic evaluation as a rst modality and therapeutic management

Endometrial Biopsy Audit and its Clinico-Pathological Correlation in Patients with Abnormal Uterine Bleeding in a Zonal Care Centre

Background: Abnormal uterine bleeding is the commonest problem affecting premenopausal, perimenopausal,and postmenopausal females and is the most common cause of the premalignant and malignant endometriallesions resulting in endometrial samples.Aim &amp; Objective: To study the clinicopathological correlation of abnormal uterine bleeding in premenopausal,perimenopausal, and postmenopausal female.Material and methods: The retrospective study was conducted in the Military Hospital, Allahabad (UP), by theDepartment of Pathology from Aug 2021 to Aug 2022. A total of 135 patients who presented with abnormal uterinebleeding were studied.Results: The age of the patients ranged from 20 to 70 years in the study. There were a total of 63 cases in the agegroup of 41 to 50 years. The major bleeding pattern observed was menorrhagia, followed by post menopausalbleeding, polymenorrhagia, metrorrhagia, oligomenorrhagia, and menometrorrhagia. The histopathologicalpatterns observed in abnormal uterine bleeding were proliferative phase endometrium, followed by secretaryphase, disordered proliferative phase, endometrial hyperplasia, atropic endometrium, malignancy, andinadequate.Conclusion: Endometrial biopsies are the first line diagnostic tool for diagnosing endometrial pathology becauseof its diagnostic accuracy, safety, quickness, and convenience. But lesion such as leiomyoma and adenomyosiscan be missed..

Single-Cell Transcriptomics of Proliferative Phase Endometrium: Systems Analysis of Cell-Cell Communication Network Using CellChat.

The endometrium thickness increases by which endometrial angiogenesis occurs in parallel with the rapid growth of endometrium during the proliferative phase, which is orchestrated by complex cell–cell interactions and cytokine networks. However, the intercellular communication has not been fully delineated. In the present work, we studied the cell–cell interactome among cells of human proliferative phase endometrium using single-cell transcriptomics. The transcriptomes of 33,240 primary endometrial cells were profiled at single-cell resolution. CellChat was used to infer the cell–cell interactome by assessing the gene expression of receptor–ligand pairs across cell types. In total, nine cell types and 88 functionally related signaling pathways were found. Among them, growth factors and angiogenic factor signaling pathways, including EGF, FGF, IGF, PDGF, TGFb, VEGF, ANGPT, and ANGPTL that are highly associated with endometrial growth, were further analyzed and verified. The results showed that stromal cells and proliferating stromal cells represented cell–cell interaction hubs with a large number of EGF, PDGF incoming signals, and FGF outgoing signals. Endothelial cells exhibited cell–cell interaction hubs with a plenty of VEGF, TGFb incoming signals, and ANGPT outgoing signals. Unciliated epithelial cells, ciliated epithelial cells, and macrophages exhibited cell–cell interaction hubs with substantial EGF outgoing signals. Ciliated epithelial cells represented cell–cell interaction hubs with a large number of IGF and TGFb incoming signals. Smooth muscle cells represented lots of PDGF incoming signals and ANGPT and ANGPTL outgoing signals. This study deconvoluted complex intercellular communications at the single-cell level and predicted meaningful biological discoveries, which deepened the understanding of communications among endometrial cells.

P-413 Non-autologous human platelet lysate stimulates in vitro proliferation of primary human endometrial cells from patients with recurrent implantation failure

Abstract Study question Does patient diagnosis impact induction of primary human endometrial cell proliferation by commercial non-autologous human platelet lysate (HPL)? Summary answer HPL stimulates in vitro proliferation of both primary endometrial epithelial cells (EECs) and stromal cells (ESCs) from patients with and without recurrent implantation failure (RIF). What is known already Inadequate endometrial receptivity and thickness are major causes for RIF. Our preliminary results suggested that non-autologous HPL stimulates in vitro proliferation of primary human EECs and ESCs isolated from patients with a history of RIF, with and without a thin endometrium (TE). In addition, using an in vitro model of embryo attachment, we reported that 48-hour treatment with HPL significantly augments the attachment of trophoblast spheroids (generated with HTR-8/SVneo cells) to EECs, suggesting an increase in endometrial receptivity following treatment. This suggests that HPL may standardize future clinical treatments for a TE and endometrial origins of RIF. Study design, size, duration Endometrial tissue was collected from five patients without RIF (control) and eighteen RIF patients at the CReATe Fertility Centre, Toronto, Canada. The eighteen RIF patients were further classified into three groups (N = 6 each) based on endometrial phenotype: 1) proliferative phase RIF only (without a TE), 2) secretory phase RIF only (without a TE), and 3) secretory phase RIF+TE (with a TE). Primary EECs and ESCs were enzymatically isolated and cultured separately. Participants/materials, setting, methods Primary EECs and ESCs were serum-starved with serum-free culture media (SFM) for 24 hours and then treated for 48 hours with the following treatment media: SFM (negative control), or SFM supplemented with 1% HPL for EECs, or 10% HPL for ESCs. Cell viability and proliferation were assessed using the metabolic assay PrestoBlue reagent and immunocytochemistry to quantify cells actively expressing the nuclear proliferation marker Ki67. Main results and the role of chance The metabolic assay demonstrated that 48-hour treatment with non-autologous HPL stimulates a similar significant increase in EEC viability and proliferation for all patient groups. EECs from patients without RIF (control) had the highest fold increase (1.49-fold, P &amp;lt;0.001), followed by proliferative phase RIF only (1.41-fold, P &amp;lt;0.001), secretory phase RIF+TE (1.39-fold, P &amp;lt;0.001), and secretory phase RIF only (1.24-fold, P &amp;lt;0.05). For ESCs, HPL stimulated a significant 2.69-fold increase in cell viability and proliferation for the secretory phase RIF+TE patient group (P &amp;lt;0.01). Pairwise comparison of the ratio of actively proliferating (Ki67+) cells between SFM and HPL treatment revealed that EECs isolated from control patients had the highest and most significant increase by 27.3% (P &amp;lt;0.05) after HPL treatment. Although not statistically significant, EECs isolated from all RIF patients displayed a similar trend of increased Ki67+ cells after HPL treatment. Whereas HPL treatment significantly increased the ratio of Ki67+ cells for ESCs isolated from both the proliferative phase (by 23.9%, P &amp;lt;0.05) and secretory phase (by 29.9%, P &amp;lt;0.05) endometrium of RIF only patients. Limitations, reasons for caution Although our data suggests that HPL treatment significantly stimulates primary endometrial cell proliferation in vitro, our sample size per group is small (N = 5-6). A larger sample size and future randomized controlled trials are needed to determine the efficacy of HPL as a treatment for a TE and/or RIF. Wider implications of the findings Our study provides the groundwork to improve clinical treatment of a TE and endometrial origins of RIF. We anticipate that in addition to stimulating cell proliferation, HPL will also induce a broad transcriptional response towards improved endometrial receptivity. We will next focus on characterizing the transcriptomic profile following HPL treatment. Trial registration number not applicable

Platelet-derived growth factor BB is reduced in endometrial endothelial cells of women with abnormal uterine bleeding–endometrial disorder

Research questionWhat is the expression pattern of platelet-derived growth factor BB (PDGF-BB), and its receptors, across the menstrual cycle in healthy control women and those with abnormal uterine bleeding–endometrial disorder (AUB-E)? DesignImmunohistochemical staining for PDGF-BB, platelet-derived growth factor receptor alpha (PDGFRα) and platelet-derived growth factor beta (PDGFRβ) was performed in control and AUB-E endometrium from the proliferative, early, mid- and late secretory phases of the menstrual cycle (n = 5 each group). Control proliferative phase endometrium was cultured in PDGF-BB (0, 10 ng/ml) and vascular maturation assessed (n = 3). Endothelial cell to vascular smooth muscle cell (VSMC) association was assessed after treatment with PDGF-BB (0, 1, 10 ng/ml). Secretion of angiogenic growth factors by endothelial cells or VSMC was determined. ResultsEndothelial cell immunoreactivity for PDGF-BB was reduced in the mid and late secretory phases in AUB-E (P = 0.008). PDGFRα was also reduced in mid secretory phase endothelial cells, proliferative and early secretory phase glandular epithelium in AUB-E (P = 0.008). PDGFRβ expression was not altered. Treatment of proliferative phase endometrium with PDGF-BB (10 ng/ml) reduced the percentage of vessels expressing contractile VSMC markers. PDGF-BB had no effect on angiogenic growth factor secretion by endothelial cells or VSMC in vitro and did not affect their association in an in-vitro endothelial cell–VSMC association assay. ConclusionsReduced endothelial cell expression of PDGF-BB in the AUB-E endometrium may contribute to the reduced vascular maturation previously observed in these women.

Histopathological pattern of endometrial biopsies in patients with abnormal uterine bleeding

Introduction: Abnormal uterine bleeding (AUB) is bleeding from the uterus that is longer than usual or that occurs at an irregular time. Bleeding may be heavier or lighter than usual and may occur often or randomly. In most women, AUB is caused by a hormone imbalance. Any woman over the age of 40 years with menorrhagia or any other forms of abnormal vaginal bleeding should undergo a gynaecological check and an endometrial biopsy as indicated, in order to detect a potentially treatable cause as atypical endometrial hyperplasia and early endometrial carcinoma. The present study was done on women with AUB to evaluate the histopathological patterns in endometrial biopsy among different age groups. Materials and Methods: A cross-sectional study was carried out in Department of Pathology, Dr. D.Y. Patil Medical College, Hospital and Research Centre, Pimpri, Pune. The clinical history and findings of 100 patients were collected and recorded. Histopathological study of endometrial patterns and age specific correlation was done. Results: Out of a total of 100 patients of AUB, 72 of them had a functional cause and the remaining 28 patients revealed an organic cause. The mean affected age was 40 years with youngest being 21 years old and the oldest patient was 57 years of age. Functional causes constituted 72% and organic lesions were seen in 28% out of which Proliferative phase endometrium was the most common functional lesion observed while endometrial hyperplasia was the commonest organic pathology seen. P value was calculated as <0.008 which was significant using chi square for trend seen in age. Conclusion: Histopathological examination of endometrial biopsy in patients of AUB is considered as a gold standard of patient evaluation, diagnosis and management and avoids any future complications.

C/EBP-β and SIRT1 regulate IL-18 expression in the proliferative phase endometria of polycystic ovary syndrome patients

BackgroundPrevious studies have shown that polycystic ovary syndrome (PCOS) patients suffer from low-grade chronic inflammation. Furthermore, our previous studies have confirmed that IL-18 is highly expressed in the serum and endometria of PCOS patients. Nevertheless, the mechanisms underlying IL-18 elevation remain unclear. This study aims to explore the signaling pathways that lead to IL-18 upregulation in the endometria of PCOS patients. MethodsUsing the TF-Search tool, we predicted that C/EBP-β may be a transcription factor for IL-18 and that deacetylase SIRT1 could be involved in its regulation. Subsequently, we analyzed SIRT1 and C/EBP-β in the proliferative endometria of PCOS patients and healthy control using immunohistochemistry, real-time quantitative PCR and western blot; PCOS diagnosis was based on the 2003 Rotterdam ESHRE/ASRM criteria. We further verified the interaction between C/EBP-β and IL-18 using a double luciferase assay. ResultsSIRT1 and C/EBP-β gene and protein levels in the proliferative endometria of PCOS patients were significantly higher than the control group. Immunohistochemical experiments confirmed that SIRT1 was primarily expressed in the endometrial nucleus, while C/EBP-β was predominantly expressed in the endometrial nucleus and cytoplasm. Double luciferase assay verified the interaction between C/EBP-β and IL-18. ConclusionSIRT1 and C/EBP-β are probable IL-18 regulatory mechanism in the endometria of PCOS patients.

Expression of BCL-2 and KI-67 in Cyclical Endometrium and in Endometrial Hyperplasia

The Present study attempts to determine the relationship between proliferation and the inhibition of apoptosis in normal endometrium and hyperplasia, using monoclonal antibodies against the proliferation marker, Ki-67 and the anti-apoptotic protein, Bcl-2 and to determine the expression of B-cell lymphoma 2 (Bcl – 2) and Ki – 67 in cyclical endometrium in proliferative and secretory phase. The Present study includes the expression of Bcl – 2 and Ki – 67 in endometrial hyperplasia.

Assessing the Efficacy of Pipelle Sampling as Outpatient Diagnostic Test in a Tertiary Care Hospital-Rawalpindi, 2020

ABSTRACT&#x0D; Objective:&#x0D; To assess the effectiveness and enlist clinical factors, which may affect effectiveness of Pipelle sampling in diagnosing endometrial pathology in patients with abnormal uterine bleeding and Post-menopausal Bleeding.&#x0D; Study Design:&#x0D; ‘Analytical Cross-Sectional’ study.&#x0D; Place and duration:&#x0D; Gynecology Department at Combined Military Hospital Rawalpindi, in collaboration with Histo-Pathology Department, of six months duration i.e. from 1st December 2019 to 31st May 2020.&#x0D; Patients and methods:&#x0D; 100 female patients of outpatient department with presenting complaints of either abnormal or post-menopausal bleeding were included in the study in the specified time frame. After informed written consent, Pipelle endometrial sampling was done on outpatient basis, and by consecutive sampling technique using lottery method, sample was sent for histopathological evaluation. Evaluated factors studied were: indications for the procedure, age, parity, age of menarche.&#x0D; Results:&#x0D; Of the total patients evaluated, 91% (n=100) were non-malignant, while 4% (n=100) were malignant. Inadequate tissue was obtained in 5% samples. Most common histopathological findings among reproductive age group were Proliferative and Secretory phase endometrium 64% (n=100). While in Post-menopausal age group 4% (n=100) were Endometrial carcinoma on histopathology. Inadequate tissue samples 5% (n=100) were noted among the Post-menopausal group. 6% samples showed endometrial hyperplasia. Pipelle Endometrial Sampling had sensitivity and specificity, in diagnosing the endometrial pathologies is summarized in Table 3.&#x0D; Conclusion:&#x0D; Pipelle Endometrial Sampling is an effective, safe, simple and acceptable procedure for diagnosing endometrial pathology. It is cost effective requiring no anesthesia with high sensitivity and specificity for detecting endometrial pathology.&#x0D;

Abnormal uterine bleeding: A study of its clinical spectrum and incidence of histopathological patterns of endometrium as a function of age with parity

Introduction: Abnormal uterine bleeding (AUB) is a common complaint affecting majority of pre and postmenopausal women. Endometirum is a dynamic tissue and is affected by a wide range of histopathological lesions. The causes of AUB is a function of age. Aim and Objective: to study the histopathological pattern in cases of AUB in reference age, parity and bleeding pattern. Patients were categorized into reproductive (50 years). Materials and Methods: A total of three hundred patients presenting with AUB are included in this study. Observation and Results: In this study patient ranged from 20 to 70 years and the mean age of the cohort was 43.46±8.07. Major parity category noted in the cohort was para-2 in 89 patients (29.67%). Major bleeding pattern observed in the cohort was menorrhagia (32.33%); followed by metrorrhagia (19.67%), menometrorrhagia (16.33%), postmenopausal bleeding (12.67%), polymenorrhoea (9.67%), polymenorrhagia (5%) and continuous bleeding (1%). Histopathological pattern observed in AUB patients was proliferative phase endometrium (31.33%), secretory phase endometrium (21.67%), menstrual endometrium (1.33%). atrophic endometrium (6%), disordered proliferation (3.33%), mixed pattern (1%), polyp (10.67%), metaplasia (9.67%), hyperplasia (8.67%), endometritis (2.67%), adenocarcinoma (1.67%) and insufficient (2%). Conclusion: The causes of AUB are function of age; perimenopausal age group is the most common affected. Dilatation & Curettage is cost effective, useful procedure but organic lesions such as leiomyoma and adenomyosis can be missed. AUB is a common disorder affecting females and is a challenge to the treating gynaecologist. Keywords: Endometrium, Premenopausal, Postmenopausal, Hyperplasia, Polyp.

Spectrum of Endometrial lesions in patients presenting with abnormal uterine bleeding

Introduction: Most of the patients with bleeding per vaginum undergo either colposcopic evaluation or Pap smear for cervical pathology or Trans Vaginal Ultrasound and endometrial biopsy to diagnose the definite pathology affecting endometrium. This study was done to see the pattern of histopathological spectrum of endometrial biopsy and their clinical correlation in a tertiary care hospital. Materials and Methods: This was a retro-prospective study conducted at a tertiary care hospital in western Maharashtra. 1000 biopsy cases were studied. Light microscopy examination and clinical diagnoses were correlated. Results: out of 1000 patient’s maximum number belonged to perimenopausal age group followed by reproductive age group and then postmenopausal age group. In 483 cases normal histopathological pattern was observed, out of which 259 cases showed proliferative phase endometrium (25.9%) and 224 cases showed secretory phase (22.4%). 104 cases showed simple (10.4%) and 43 cases (4.3%) complex hyperplasia. Exogenous hormonal effect was seen in 148 cases (14.8%). The least pattern which was observed was endometrial carcinoma, seen in only 2.3% of patients. Conclusion: The study showed that carcinoma was most common in the post-menopausal age group. This group also had the maximum frequency of atrophic endometrium and complex hyperplasia. Proliferative endometrium was recorded highest in perimenopausal age group while reproductive age group showed highest frequency of secretory type endometrium. This study is in concordance with most other conducted studies on similar parameters. Keywords: Biopsy, Endometrium, Uterine bleeding.

Histopathological spectrum of endometrial lesions in cases of abnormal uterine bleeding at a Tertiary Care Hospital.

Objectives: To assess histological spectrum of endometrial lesions in abnormal uterine bleeding at a tertiary care hospital. Study Design: Cross Sectional study. Setting: Department of Pathology, Multan Medical &amp; Dental College, Multan and Faisalabad Medical University, Faisalabad. Period: January, 2019 to December, 2019. Material &amp; Methods: Using non – probability purposive sampling technique. Results: A total of 238 ladies presenting with abnormal uterine bleeding were registered in our study. Mean age of these patients with AUB was 47.25 ± 5.57 years (ranging 21 years to 59 years), 158 (66.4%) were aged more than 45 years. Eighty five (35.7%) were from rural areas while 153 (64.3%) from urban areas. Of these 238 females, 162 (68.1%) were illiterate and 76 (31.9%) were literate. Majority of patients i.e. 218 (91.6%) were married and mean duration of illness was 5.28 ± 3.89 months while 169 (71%) presented within 6 months of illness. Histopathological pattern revealed Proliferative phase endometrium was observed in 69 (29%), endometrial hyperplasia in 59 (24.8%), chronic endometritis in 40 (16.8%), secretory phase in 40 (16.8 %), atrophic endometrium in 30 (12.6%) and endometrial polyps in 10 (4.2%). Conclusion: Histological pattern revealed proliferative endometrium was commonest followed by hyperplasia, secretory phase and chronic endometrium in our study. Disordered proliferative endometrium was commonly observed in married females and those having duration of symptoms more than 6 months. Hyperplasia was more frequent in older patients who were married. Secretory phase was significantly more common in patients with prolonged disease and chronic endometritis was associated with marital status and prolonged disease duration.